A new hybrid protein is a prominent player reducing cytokine storm
caused by excessively activated macrophages.
Abstract
We newly developed a hybrid protein, tentatively named human MIKO-1
(hM1), based on the amino acid sequences of human S100A8 (hS100A8) and
hS100A9. Human macrophage-like cells (hMΦ), which were differentiated
from THP-1 cells by phorbol 12-myristate 13-acetate, were used to
investigate the immune function of hM1 and its dynamic mobility in
cells. Western blotting was conducted to detect hM1 in the cytoplasm and
nucleus of hMΦ. A polymerase chain reaction (PCR) and quantitative PCR
were performed to examine changes in the mRNA levels of proinflammatory
cytokines in hMΦ. Fluorescent immunochemical staining was conducted to
microscopically observe the intracellular behavior of hM1 in hMΦ,
together with hS100A8 and/or hS100A9. Microscopic observations showed
that hM1 rapidly bound to activated hMΦ and immediately migrated even
inside the nucleus of these cells. The intracellular localization of hM1
in hMΦ almost coincided with that of hS100A8, suggesting the close
involvement of hS100A8 in the intracellular behavior of hM1 in hMΦ. The
mRNA expression of each proinflammatory cytokine was significantly
suppressed in hMΦ preliminarily treated with hM1 despite a subsequent
stimulation with lipopolysaccharide. Significant decreases in the mRNA
levels of proinflammatory cytokines in hMΦ strongly suggest the
potential of hM1 as a negative regulator of inflammation. hM1 may land
on a special site on the DNA chain, such as the NF-κB motif, in the
nucleus to directly inhibit the mRNA expression of proinflammatory
cytokines. In conclusion, hM1 is a prominent player that significantly
attenuates the cytokine storm caused by excessively activated hMΦ.